Journal: International Journal of Molecular Sciences

Article Title: Evaluating Whether Radiofrequency Irradiation Attenuated UV-B-Induced Skin Pigmentation by Increasing Melanosomal Autophagy and Decreasing Melanin Synthesis

doi: 10.3390/ijms221910724

Figure Lengend Snippet: Reduction in melanin accumulation via autophagy activation by RF irradiation. ( A ) Pigmentation changes in UV-B-exposed mouse skin were assessed 1, 7, and 28 days after RF irradiation (upper row). Melanin content was assessed by FM staining (lower row; scale bar = 100 μm); ( B , C ) Quantitative graphs of representative mouse skin images and ( D , E ) representative FM staining images; ( F ) HEMn were treated either with α-MSH (200 μM) or with α-MSH/RF (2 MHz, 10 W for 100 ms) in vitro, or otherwise not irradiated (control); ( G , H ) The mRNA expression levels of MC1R ( F ), and MITF ( G ) were identified in the skin tissue. All mRNA levels were measured by qRT-PCR, normalized versus ACTB, and expressed relative to levels in the control; ( I ) Melanin content was measured in HEMn after RF irradiation; ( J ) Changes in melanin after RF irradiation were observed with TEM. Data are presented as mean ± SD. **, p < 0.01, vs. control 28 days (skin) or control (HEMn); $, p < 0.05; $$, p < 0.01, vs. UV-B 28 days (Skin) or α-MSH (HEMn); †, p < 0.05; ††, p < 0.01, vs. 1 day after RF irradiation (Mann–Whitney U test). α-MSH, alpha-melanotropin; FM, Fontana-Masson staining; HEMn, human epidermal melanocyte; NF- κ B, nuclear factor kappa-light-chain-enhancer of activated B cells; MC1R, melanocortin 1 receptor; MITF, micropthalmia-associated transcription factor; RF, radiofrequency; TEM, transmission electron microscopy; TNFR, tumor necrosis factor receptor; TLR4, Toll-like receptor 4; UV-B, ultraviolet-B; UV-B/RF, UV-B plus RF.

Article Snippet: The skin tissues were incubated in Fontana ammoniacal silver solution (ScyTek, West Logan, UT, USA) overnight at room temperature, subsequently rinsed three times with distilled water, and then incubated in hypo solution for 3 min.

Techniques: Activation Assay, Irradiation, Staining, In Vitro, Control, Expressing, Quantitative RT-PCR, MANN-WHITNEY, Transmission Assay, Electron Microscopy

Journal: Scientific Reports

Article Title: Beauvericin inhibits melanogenesis by regulating cAMP/PKA/CREB and LXR-α/p38 MAPK–mediated pathways

doi: 10.1038/s41598-018-33352-8

Figure Lengend Snippet: The inhibitory effect of beauvericn on melanogenesis in human epidermal melanocytes. ( A ) Human epidermal melanocytes were treated with beauvericin for 48 h. After harvesting, the cells were dissolved in a mixture of Soluene-350 and water. The melanin content was measured by absorbance at 500 nm. P -values were obtained by the one-way ANOVA. ( B ) Human epidermal melanocytes were treated with beauvericin for 48 h. After harvesting, the cells were lysed by sonication and assayed for cellular tyrosinase activity (dopa oxidase). Absorbance was immediately measured at 505 nm. Results were confirmed from at least three independent experiments, and values represent the means ± SEM. * P < 0.05 vs. untreated control. P -values were obtained by the one-way ANOVA. ( C ) Cell counting kit-8 was used to assay cell viability. Results were confirmed from at least three independent experiments, and values represent the means ± SEM. * P < 0.05 versus untreated control. P -values were obtained by the one-way ANOVA. ( D ) Reconstructed epidermis was incubated with beauvericin (10 μM) for 3 days. The epidermis was subjected to Fontana–Masson staining. Quantification of Fontana-Masson staining was performed using Image J. Data are presented as the means ± SEM of three independent experiments. * P < 0.05 vs. control group. P -values were obtained by Student’s t -test. ( E ) After B16F10 cells and human epidermal melanocytes were incubated with the indicated concentration of beauvericin for 24 h or 48 h, imaging analysis was performed. Fk, forskolin; BEA, beauvericin.

Article Snippet: Briefly, the epidermal cells were washed twice with dH 2 O and then incubated with Fontana ammoniacal silver solution (American MasterTech, Lodi, CA, USA) at 37 °C for 1 h. For staining of the reconstructed epidermis, Fontana-Masson Staining Kit (American MasterTech) was used according to the manufacturer’s instructions.

Techniques: Sonication, Activity Assay, Control, Cell Counting, Incubation, Staining, Concentration Assay, Imaging